You synthesized an enhancerless GFP reporter gene in which the GFP cDNA is placed downstream of a promoter that functions in mice. You have fused this enhancerless GFP reporter to fragments of genomic DNA surrounding the transcribed part of a mouse gene. Each reporter construct is introduced into the nucleus of a fertilized mouse egg (zygote) to make transgenic mice. You analyze the pattern of GFP expression in these mice and obtain the following results. Genomic DNA surrounding a mouse gene 1 2 3 GFP expression pattern GFP Eye and Leg GFP Eye GFP Leg GFP Eye and Leg PAX6 gene regulates development of the eye across many species. You want to test if the human PAX6 protein can substitute the mouse PAX6 protein in eye development. What sequence do you need to fuse to the human PAX6 cDNA to cause PAX6 expression in mouse eyes? Mouse promoter Mouse promoter and region 3 Mouse promoter and region 2 Random mouse genome sequences Mouse promoter and region 1 None because human regulatory sequences are needed
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You synthesized an enhancerless GFP reporter gene in which the GFP cDNA is placed downstream of a promoter that functions in mice. You have fused this enhancerless GFP reporter to fragments of genomic DNA surrounding the transcribed part of a mouse gene. Each reporter construct is introduced into the nucleus of a fertilized mouse egg (zygote) to make transgenic mice. You analyze the pattern of GFP expression in these mice and obtain the following results: Genomic DNA surrounding a mouse gene 1 2 3 GFP expression pattern Eye and Leg Eye Leg Eye and Leg The region containing an eye-specific enhancer of the mouse gene is 1. The region that is NOT required for either eye or leg expression is most likely [Select].
Josee P.
3. You have synthesized and enhancerless GFP reporter gene in which the jellyfish GFP cDNA is placed downstream of a basal promoter that functions in mice. You will now fuse this enhancerless reporter to three types of sequences listed below (i-iii). a. Which of the three types of sequences would you use for which of the three listed purposes? In each case, explain how the particular fusion would address the particular use. Types of sequences fused to the reporter: i. Random mouse genome sequences ii. Known mouse kidney-specific enhancer iii. Fragments of genomic DNA surrounding the transcribed part of a mouse gene Uses: 1) To identify a gene's enhancer(s) 2) To express GFP tissue-specifically 3) To identify genes expressed in neurons b. Which of the sequences (i-iii) would you fuse to a particular mouse gene of interest in order to express the protein product of the gene ectopically, that is, in a tissue in which the gene is not usually expressed? Why might you want to do this experiment in the first place?
You have synthesized an enhancerless GFP reporter gene in which the jellyfish $G F P$ cDNA is placed downstream of a basal promoter that functions in mice. You will now fuse this enhancerless reporter to the three types of sequences listed below (x-z). a. Which of the three types of sequences would you use for which of the three listed purposes (i-iii)? In each case, explain how the particular fusion would address the particular use. Types of sequences fused to the reporter: x. random mouse genome sequences y. known mouse kidney-specific enhancer z. fragments of genomic DNA surrounding the transcribed part of a mouse gene Uses: i. to identify a gene's enhancer(s) ii. to express GFP tissue-specifically iii. to identify genes expressed in neurons b. Which of the sequences $(x-z)$ would you fuse to a particular mouse gene of interest in order to express the protein product of the gene ectopically, that is, in a tissue in which the gene is not usually expressed? Why might you want to do this experiment in the first place?
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